Nucleic acid sequence measuring method, nucleic acid sequence measuring device, manufacturing method for nucleic acid sequence measuring device, and nucleic acid sequence measuring apparatus
First Claim
1. A nucleic acid sequence measuring method comprising:
- preparing a sample solution containing a target nucleic acid;
supplying the sample solution to a nucleic acid sequence measuring device; and
measuring fluorescence from the nucleic acid sequence measuring device, the nucleic acid sequence measuring device including;
a fluorescent probe having a coupling part and a base end and added with a fluorescent molecule at a predetermined position;
a quenching probe having a coupling part and a base end and added with a quenching substance at a predetermined position; and
a substrate having a solid surface to which the base end of each of the fluorescent probe and the quenching probe is fixed, wherein;
the coupling part of the fluorescent probe and the coupling part of the quenching probe each have a mutually complementary nucleic acid sequence;
at least one of the fluorescent probe and the quenching probe has a detection part with a nucleic acid sequence complementary with a particular nucleic acid sequence; and
the base end of the fluorescent probe and the base end of the quenching probe are fixed to the solid surface, the probes are in such a positional relationship that fluorescence from the fluorescent molecule is quenched by the quenching substance that has approached the fluorescent molecule by maintaining the coupling between the fluorescent probe and the quenching probe through the coupling parts when the hybridization between the target nucleic acid and the detection part has not occurred, and that fluorescence is emitted from the fluorescent molecule separated from the quenching substance due to canceling of the coupling between the fluorescent probe and the quenching probe through the coupling parts when the hybridization between the target nucleic acid and the detection part has occurred.
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Abstract
A nucleic acid sequence measuring method includes measuring fluorescence from the nucleic acid sequence measuring device supplied with a sample solution. The device includes a fluorescent probe added with a fluorescent molecule, and a quenching probe added with a quenching substance. The fluorescent probe and/or the quenching probe has a detection part detecting a predetermined nucleic acid sequence. Fluorescence from the fluorescent molecule is quenched by the quenching substance coupled with the fluorescent molecule when the hybridization between the detection target nucleic acid and the detection part has not occurred, and fluorescence is emitted from the fluorescent molecule separated from the quenching substance when the hybridization has occurred.
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Citations
16 Claims
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1. A nucleic acid sequence measuring method comprising:
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preparing a sample solution containing a target nucleic acid; supplying the sample solution to a nucleic acid sequence measuring device; and measuring fluorescence from the nucleic acid sequence measuring device, the nucleic acid sequence measuring device including; a fluorescent probe having a coupling part and a base end and added with a fluorescent molecule at a predetermined position; a quenching probe having a coupling part and a base end and added with a quenching substance at a predetermined position; and a substrate having a solid surface to which the base end of each of the fluorescent probe and the quenching probe is fixed, wherein; the coupling part of the fluorescent probe and the coupling part of the quenching probe each have a mutually complementary nucleic acid sequence; at least one of the fluorescent probe and the quenching probe has a detection part with a nucleic acid sequence complementary with a particular nucleic acid sequence; and the base end of the fluorescent probe and the base end of the quenching probe are fixed to the solid surface, the probes are in such a positional relationship that fluorescence from the fluorescent molecule is quenched by the quenching substance that has approached the fluorescent molecule by maintaining the coupling between the fluorescent probe and the quenching probe through the coupling parts when the hybridization between the target nucleic acid and the detection part has not occurred, and that fluorescence is emitted from the fluorescent molecule separated from the quenching substance due to canceling of the coupling between the fluorescent probe and the quenching probe through the coupling parts when the hybridization between the target nucleic acid and the detection part has occurred. - View Dependent Claims (2)
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3. A nucleic acid sequence measuring device configured to measure a predetermined nucleic acid sequence of a target nucleic acid in a sample solution, comprising:
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a fluorescent probe having a coupling part and a base end and added with a fluorescent molecule at a predetermined position; a quenching probe having a coupling part and a base end and added with a quenching substance at a predetermined position; and a substrate having a solid surface to which the base end of each of the fluorescent probe and the quenching probe is fixed, wherein; the coupling part of the fluorescent probe and the coupling part of the quenching probe each have a mutually complementary nucleic acid sequence; at least one of the fluorescent probe and the quenching probe has a detection part with a nucleic acid sequence complementary with a particular nucleic acid sequence; and the base end of the fluorescent probe and the base end of the quenching probe are fixed to the solid surface, the probes are in such a positional relationship that fluorescence from the fluorescent molecule is quenched by the quenching substance that has approached the fluorescent molecule by maintaining the coupling between the fluorescent probe and the quenching probe through the coupling parts when the hybridization between the target nucleic acid and the detection part has not occurred, and that fluorescence is emitted from the fluorescent molecule separated from the quenching substance due to canceling of the coupling between the fluorescent probe and the quenching probe through the coupling parts when the hybridization between the target nucleic acid and the detection part has occurred. - View Dependent Claims (4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16)
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Specification